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AR226-3206 TRADE SECRET Study Title H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Supplement No. 1 Laboratory Project ID: DuPont-2931 AUTHOR: Carol Pinlay, B.A. STUDY COMPLETED ON: August 29, 2001 SUPPLEMENT NO. 1 COMPLETED ON; December 12,2001 PERFORMING LABORATORY: E.I. du Font de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 WORK REQUEST NUMBER--:Mm1j SERVICE CODE NUMBER:--------|f Page 1 of 40 Company Sanitized. Does rot contain TSCA CBI H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2931 Supplement No. 1 CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study. Renewed by; yi^cl^t^ \J!jtjOUC(^ C. Judith C. ^- Stadler, Ph.D., D.A.B.T. Diceetor ^->GQ^Od/ Date /L/L^ Issued by Study Director: ______C Cat^Fsp^ayu,Bk/.jAO.^^.______ Staff Scientist f'l -/^^^ ^Z> 6| Date -2- Company Sanitized. Does not contain TSCA CBI TABLE OF CONTENTS Page CERTIFICATION........................................................................................................................^ LIST OF TABLES .........................................................................................................................4 LIST OF FIGURES.......................................i........................>.........."4 LIST OF APPENDICES...............................................................................................................4 STUDY INFORMATION ...........................................................................................................5 STUDY PERSONNEL...............................................................................................................-^ REASON FOR SUPPLEMENT NO. 1 ........................................................................................8 SUMMARY..............................................................-...--....--.......................................................9 MATERIALS AND METHODS ................................................................................................10 RESULTS AND DISCUSSION ..................................................................................................10 A. Liver Weights....................................................................................................................10 1. Test Substance.......................... .......................................................................-.....---.--- ............10 2. Positive Controls....................................................................................................................................10 B. Liver Fluorine Data...........................--..............................................................................11 1. Positive Controls..................................--------.................--......................------..............--..................11 2. TestSubstance.................................................................................--.----------------ll CONCLUSIONS..................................................................................--"."--...........................11 "- | A' KJ jr^^^^-.-.-.^..-.-.....-.-.B--.a...-a.B.flM..BBP*>*>***"********"-I^ FIGURES ..........,,..............................................................................................."...."--.........16 APPENDICES..............................................................................................................................22 -3- Company Sanitized. Does not contain TSCA CBI H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2931 Supplement No. 1 LIST OF TABLES Page 1. MEAN BODY AND LIVER WEIGHTS .........................................................................................................13 2. MEAN LIVER FLUORINE LEVELS......................................................--....................................................... 15 3. MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE..................................................15 LIST OF FIGURES Page 1. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND NEGATIVE 17 CONTROLS.......................................................................................................................--.............................. 2. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND POSITIVE CONTROLS....................................................................................................................................................... -18 3. COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTRATION FOR H-24042 AND NEGATIVE CONTROLS.........................................--............--.--............................19 4. MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE.......--.......................---- -20 5. COMPARISON OF MEAN BLOOD AND MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE........................................--...........----.....--......----...------...................... --21 LIST OF APPENDICES Page A. INDIVIDUAL FINAL BODY WEIGHTS, LIVER WEIGHTS, AND RELATIVE LIVER TO BODY WEIGHT--...........................................----.......--...................-.......--..................................--.--. .........23 B. INDIVIDUALFLUORINELEVELS............................................................--.......----....--.--..........---- ..34 -4- Company Sanitized. Does not contain TSCA CBi H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats STUDY INFORMATION TEST SUBSTANCE: Substance Tes Haskell Number: 24042 Compositi DuPont-2931 Supplement No. 1 Known Impurities: Poly(difluoromethylene), alpha-fluoro-omega-(2-iodoethyl)(2-3%) Physical Characteristics: Pale yellow to amber brown waxy solid POSITIVE CONTROL: Substance Tested! Synonyms/Codes: Haskell Number: 24019 -5- Company Sanitized. Does not contain TSCA CBI Known Impurities: Unknown POSITIVE CONTROL: Known Impurities: Unknown Sponsor: E. I. du Font de Nemours and Company Wilmington, Delaware 19898 U.S.A. Study Initiated/Completed: April 30,1999 / (see report cover page) In-Life Initiated/Completed: June 13,1999 / September 14,1999 -6- Gompany Sanitized. Does not contain TSCACBI H-24042: Biopersistence Screening 10-Dose Oral Gavage Stady in Rats STUDY PERSONNEL Study Director: Management: Primary Technician: Carol Finlay, B.A. Judith C. Stadler, Ph.D., D.A.B.T. Richard P. Mathena liver Fluorine Data Analysis: Paul M. Hinderiiter, Ph.D. Management: Matthew S. Bogdanffy, Ph.D., D.A.B.T. Toxicology Report Preparation: Wanda F. Dinbokowitz Laboratory Veterinarian: Wanda L. West, D.V.M., A.C.L.A.M. DuPont-2931 Supplement No. 1 -7- Company Sanitized. Does not contain TSCA CBI H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2931 Supplement No. 1 REASON FOR SUPPLEMENT NO. 1 The purpose of this supplement is to present the results from analysis of selected liver samples from rats treated with the test substance (H-24042), 2 positive controls (H-24019 and H-24020), and 2 negative controls (corn oil and corn oil/acetone). -8- Company Sanitized. Does not contain TSCA CBI SUMMARY The objective of this study was to evaluate the potential for H-24042, when administered by gavage, to be absorbed and to accumulate in a mammalian system. Six groups of 5 male Cri:CD(SD)IGSBR rats each were given 100 mg/kg/day of H-24042. The test substance was administered to one group of 5 rats for 5 consecutive days and to 5 groups for 10 days. On selected days (5,10,13,24,52,94) 5 rats per group were euthanized and the blood and livers were collected, livers were weighed and those collected on study days 10 and 94 were analyzed for total fluorine. This supplement includes liver weight data and the results of total fluorine analyses in liver. Additionally, 2 negative controls, corn oil and corn oiVacetone (80:20), and 2 positive controls, H-24019 (10 mg/kg/day) and H-24020 (20 mg/kg/day), were tested as described for H-24042. The mean relative liver weight (liver/body weight) of rats dosed with the test substance, H-24042, was up to 39% higher at day 10 than the mean relative liver weights of the negative control groups. By day 94, the mean relative liver weight of rats dosed with the test substance was similar to the negative controls. On day 10, the mean relative liver weight of rats dosed with the positive control H-24019 was up to 27% higher than the negative controls. By day 94, the mean relative liver weight was up to 28% higher than the negative controls. In rats given H-24020, the mean relative liver weight was up to 75% higher than the negative controls. By day 94, the weights were similar. The concentration of fluorine in the livers from rats dosed with the test substance, H-24042, was 271.39 pM equivalents on day 10 and 15.15 piM equivalents on day 94. On day 10, mean [iM equivalent concentrations of fluorine in the livers from rats dosed with the positive control materials were approximately 18-fold (H-24019) and 4-fold (H-24020) greater than the fluorine concentration in livers from rats treated with the test substance. By day 94, the fluorine concentration in rats dosed with the positive control H-24019 was approximately 80x the fluorine concentration in rats treated with H-24042. The fluorine concentration in rats dosed with the positive control H-24020 and test substance H-24042 was similar by day 94. The IJI.M equivalents of fluorine in the livers from rats dosed with the test substance, H-24042, were higher than levels in the blood. Under the conditions of this study, there was some absorption and retention of fluorine in the liver from rats dosed with H-24042. Fluorine levels in the livers from rats dosed with the test substance were lower than levels in rats dosed with the positive control H-24019. Fluorine levels in the livers from rats dosed with the test substance were lower on day 10 than levels in rats dosed with the positive control H-24020. On day 94, the fluorine levels were similar. liver weights were elevated in rats dosed with the test substance, H-24042, at the end of the dosing period, but not at the end of the recovery period. -9- Company Sanitized. Does not contain TSCA CBI H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2931 Supplement No. 1 MATERIALS AND METHODS Blood and livers were collected on days 5,10,13,24, 52, and 94. The livers were weighed. Only the test day 10 and 94 livers were analyzed for total fluorine. These time points provided sufficient data to determine the accumulation and persistence of fluorine in the liver. See the original report for details of the analysis. RESULTS AND DISCUSSION A. Liver Weights (Table 1, Figures 1-3, Appendix A) As with the body weights, comparison of absolute liver weights was complicated by the fact that there was a difference in age between rats dosed with the test substance and those dosed with the H-24019 positive control and corn oil:acetone negative control. Therefore, liver weights relative to total body weight are presented for comparison for all rats. 1. Test Substance The mean relative liver weight of rats dosed with the test substance, H-24042, was up to 36% higher than the negative controls on day 10. On day 13, the mean relative liver weight of rats dosed with the test substance was up to 48% higher than the negative controls. The weights were similar throughout the remainder of the recovery period. 2. Positive Controls The mean relative liver weights of rats dosed with one of the positive controls, H-24019, were slightly lower on days 10 and 13 than the liver weights of rats dosed with the test substance, H-24042. The liver weights of rats dosed with H-24019 were 13-21% higher during the remainder of the recovery period than the liver weights of rats dosed with the test substance. The mean relative liver weight of rats dosed with H-24020, the other positive control, was 26% higher on day 10 than the liver weight of rats dosed with H-24042. On days 13 and 24 (recovery), the mean relative liver weights of rats dosed with H-24020 were 10 or 40% higher than the liver weights of rats dosed with H-24042. On days 52 and 94, the weights were similar. The mean relative liver weight of rats dosed with H-24019 was up to 27% higher at day 10 than the negative control group. By day 94, the mean relative liver weight of rats dosed with H-24019 was up to 28% higher than the negative controls. The mean relative liver weight of rats dosed with H-24020 was up to 75% higher at day 10 than the negative controls. The mean relative liver weights of rats dosed with H-24020 began to decrease during the recovery period and were similar by test day 94 to the negative controls. -10- Ccmpany Sanitized. Does not contain TSCA CBI H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2931 Supplement No. 1 Therefore, liver weights of rats dosed with the test substance, H-24042, were elevated at the end of the 10-day dosing period. The relative liver weight by the end of recovery in rats dosed with the test substance was lower than in rats dosed with the positive control material H-24019 and similar to rats dosed with the positive control material H-24020. B. Liver Fluorine Data (Tables 2 and 3, Figures 3-5, Appendix B) 1. Positive Controls The positive controls were H-24019 and H-24020. The concentrations of fluorine in the livers on day 10 from rats dosed with the positive control materials were 5045.85 and 1127.25 juM equivalents for H-24019 and H-24020, respectively. By day 94, the concentrations were 1190.15 and 10.72 ^M equivalents. 2. Test Substance Levels of total fluorine in livers from rats dosed with the test substance, H-24042, were generally lower than the levels in livers from rats dosed with the positive control materials. The total fluorine concentration in the liver from rats dosed with H-24042 was 271.39 fJiM equivalents at day 10 and 15.15 /tM equivalents at day 94. For the positive control H-24019, the liver concentrations were approximately 18x higher (day 10) and approximately 80x higher (end of study) than H-24042. For H-24020, the liver concentration was approximately 4x higher by day 10. The fluorine concentration in rats dosed with the positive control H-24020 and test substance H-24042 was similar by day 94. The fiM equivalents of fluorine in the livers from rats dosed with the test substance were higher than levels in the blood. CONCLUSIONS Under the conditions of this study, there was some absorption and retention of fluorine in the liver from rats dosed with H-24042. Fluorine levels in the livers from rats dosed with the test substance were lower than levels in rats dosed with the positive control H-24019. Fluorine levels in the livers from rats dosed with the test substance were lower on day 10 than levels in rats dosed with the positive control H-24020. On day 94, the fluorine levels were similar. Liver weights were elevated in rats dosed with the test substance, H-24042, at the end of the dosing period, but not at the end of the recovery period. Company Sanlfeed. Does not contain TSCA CBi TABLES -12- Company Sanitized. Does not contain TSCA CBI H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2931 Supplement No. 1 Test Days 5 10 13 24 52 94 TABLE 1 MEAN BODY AND UVER WEIGHTS (g) CORN OIL (NEGATIVE CONTROL) Absolute Body Weight liver Weight 259.3 12.620 311.1 15.861 322.9 14.906 391.2 16.635 464.7 17.240 557.8 20.118 Mean Relative liver Weight(% of Body Weight) 0.049 0.051 0.046 0.043 0.037 0.036 Test Days 5 10 13 24 52 94 CORN OIL:ACETONE (NEGATIVE CONTROL) Absolute Body Weight Liver Weight 313.4 13.632 356.5 15.597 377.6 17.715 423.5 17.172 527.6 21.275 567.8 18.447 Mean Relative liver Weight(% of Body Weight) 0.043 0.044 0.047 0.041 0.040 0.032 Test Days 5 10 13 24 52 94 H-24019 (POSITIVE CONTROL) Absolute________ Body Weight Liver Weight 306.6 15.456 310.1 17.336 344.8 22.049. 387.9 21.313 438.1 19.826 597.9 24.455 Mean Relative liver Weight(% of Body Weight) 0.050 0.056 0.064 0.055 0.045 0.041 -13 Company Sanitized. Does not contain TSCACB1 H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2931 Supplement No. 1 Test Days 5 10 13 24 52 94 TABLE 1 (Continued) MEAN BODY AND LIVER WEIGHTS (g) H-24020 (POSITIVE CONTROL) Absolute BodyWeigiht Liver Weight 244.8 17.052 284.4 21.944 289.1 21.780 370.3 20.619 456.7 17.713 538.7 18.205 Mean Relative Liver Weight(% of Body Weight) 0.070 0.077 0.075 0.056 0.039 0.034 Test Days 5 10 13 24 52 94 H-24042 (TEST SUBSTANCE) Absolute Body Weight Liver Weight 258.9 16.273 286.3 17.575 317.7 21.447 385.2 17.580 480.1 19.048 592.0 20.150 Mean Relative liver Weight(% of Body Weight) 0.063 0.061 0.068 0.046 0.040 0.034 -14- Comparry Sanitized. Does not contain TSCA CBE TABLE 2 MEAN LIVER FLUORINE LEVELS Test Days 10 94 Nep;aitivelControls Corn Oil (ppm) Corn ( )il:Acetone (ppm) <0.2 0.6s1 <0.5 I2 Positive Controls H-24019 (ppm) 328.18 (19.3)" 77.56 (10.1) H-24020 (ppm) 155.76 (29.2) 1.68 (0.3) a One value. Pour of the values were below the level of detection (LOD). b Standard deviation is in parentheses. Test Substance H-24042 (ppm) 203.74 (20.45) 11.56 (2.22) TABLE 3 MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE Test Days 10 94 Positive (controls H-24019 H-24020 pM. F Equivalents 5045.85 (296.91)3 /iM F Equivalents 1127.25 (211.37) 1190.15 (156.05) 10.72 (2.14) a Standard deviation is in parentheses. Test Substance H-24042 juM F Equivalents 271.39 (27.26) 15.15 (2.96) -15- Connpany Sanitized. Does not contain TSCA CBI H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2931 Supplement No. 1 FIGURES -16- Company Sanitized. Does not contain TSCA CBI H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats FIGURE 1 COMPARISON OP MEAN RELATIVE UVER WEIGHTS: TEST SUBSTANCE AND NEGATIVE CONTROLS Test Days -17- Company Sanitized. Does not contain TS H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats FIGURE 2 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND POSITIVE CONTROLS i ^ wij '~** ^ W ^- I ^ S 3 ls ^ ^ B ^i-' -18- Company Sanitized. Does not coiitam H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats FIGURES COMPARISON OF RELATIVE UVER WEIGHT AND MEAN UVER FLUORINE CONCENT FOR H-24042 AND NEGATIVE CONTROLS 2SO< 150-T Day 10 Day 13 Day 94 Day 10 1 Cora Oil (Negative Control) Corn OiLAcetone (Negative Control) 13 H-24042 (Test Substance) 19CBSS?anSxanitized. Does not contain H-24042: Biopersistence Screening, 10-Dose Oral Gavage Study in Rats FIGURE 4 MEAN LTVER FLUORINE CONCENTRATION NORMALIZED TO DOSE 6000-r 5000-h 4000-4 s 3000 te 20004 1000 Day 10 Day 94 -20- Company Sanitized. Does not contain H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats FIGURE 5 COMPARISON OF MEAN BLOOD AND MEAN UVER FLUORINE CONCENTRATION NORMALI Blood / Liver Blood /Liver Blood / Liv -21- Ccmpany Sanitized. Does not conlain H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2931 Supplement No. 1 APPENDICES Company Sanitized. Does not contain TSCACB1 H-24042: Biopersistence Screening 10-Doss Oral Gavage Study in Rats DuPont-2931 Supplement No. 1 APPENDIX A Individual Final Body Weights, Liver Weights, and Relative Liver to Body Weight -23- Company Sanitized. Does not contain TSCA CBI H-24042: Biopersistelice Screening 10-Dose Oral Gavage iStudy in Rats DuPont-2931 Supplement No. 1 CORN OIL (NEGATIVE CONTROL) TNDJ\fTDUAL FINAL BODY WEIGHTS (g) , LIVER WEIGHTS (<?). AND RELATIVE L:HFEB. TO BODY WEHGHT OF MALE RATS Animal Number 627789 627801 627805 627810 627818 Final Body Wt. 247.7 240.1 283.7 265.0 260.2 GROUP I Liver Wt. 11.428 12.216 14.081 13.395 11.981 Relative Liver to Body Wt. 0.046 0.051 0.050 0.051 0.046 Test Day 5 5 5 5 5 Animal Number 627793 627797 627799 627800 627808 Final Body Wt. 283.5 324.2 294.7 331.3 321.8 GROUP II Liver Wt. 15.292 15.803 14.285 16.344 17.580 Relative Liver to Body Wt. 0.054 0.049 0.048 0.049 0.055 Test Day 10 10 10 10 10 Animal Number 627796 627806 627807 627809 627816 Final Body Wt. 322.7 320.3 305.2 336.2 330.2 GROUP III Liver Wt. 13.404 15.413 14.935 14.967 15.811 Relative Liver to Body Wt. 0.042 0.048 0.049 0.045 0.048 Test Day 13 13 13 13 13 .24- Company Sanitized. Does not contain TSCACBI H-24042: Biopersistciice Screening 10-Dose Oral Gavage:Study in Rats DuPont-2931 Supplement No. 1 CORN OIL (NEGATIVE CONTROL) INDI\riDUAL FINAL BODY WEIGHTS (g) ,, LIVER WEIGHTS (gr), AND RELATIVE L:H7ER TO BODY WEIGHT OF MALE RATS Animal Number 627791 627794 627804 627812 627817 Final Body Wt. 365.6 416.7 378.1 388.4 407.3 GROUP IV Liver Wt. 15.371 19.105 15.326 16.775 16.600 Relative Liver to Body Wt. 0.042 0.046 0.041 0.043 0.041 Test Day 24 24 24 24 24 Animal Number 627792 627798 627813 627814 627815 Final Body Wt. 496.8 517.3 416.8 386.9 505.9 GROUP V Liver Wt. 16.359 19.293 15,234 14.358 20.958 Relative Liver to Body Wt. 0.033 0.037 0.037 0.037 0.041 Test Day 52 52 52 52 52 Animal Number 627795 627811 627819 627820 627821 Final Body Wt. 485.0 511.6 653.1 618.0 521.5 GROUP VI Liver Wt. 16.649 17.405 21.828 26.766 17.943 Relative Liver to Body Wt. 0.034 0.034 0.033 0.043 0.034 Test Day 94 94 94 94 94 -25- Company Sanitized. Doss not contain TSCA CBI H-24042: Biopersistelice Screening 10-Dose Oral Gavage,Study in Rats DuPont-2931 Supplement No. 1 CORN OILi: ACETONE (MEGAI'IVE CONTROL) IMDI\FTDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g ), AND RELATIVE LT\7ER TO BODY WEIIBHT OF MALE RATS Animal Mumber 625361 625363 625364 625380 625383 Final Body Wt. 349.7 286.7 312.0 305.5 313.1 GROUP I Liver Wt. 16.800 11.236 13.845 13.253 13.026 Relative Liver to Body Wt. 0.048 0.039 0.044 0.043 0.042 Test Day 5 5 5 5 5 Animal Number 625355 625362 625367 625368 625378 Final Body Wt. 333.6 332.9 363.8 386.7 365.4 GROUP II Liver Wt. 14.409 14.363 16.139 17.745 15.328 Relative Liver to Body Wt. 0.043 0.043 0.044 0.046 0.042 Test Day 10 10 10 10 10 Animal Number 625357 625358 625370 625384 625385 Final Body Wt. 405.6 340.7 407.0 357.1 377.6 GROUP III Liver Wt. 20.091 12.954 21.654 16.520 17.355 Relative Liver to Body Wt. 0.050 0.038 0.053 0.046 0.046 Test Day 13 13 13 13 13 -26- San,^."^"0'0""""^061 Company H-24042: Biopersistelace Screening 10-Dose Oral Gavage Study in Rats DuPont-2931 Supplement No. 1 CORN OIL^ACETONE (MEGA1;fIVE CONTROL) iinm/TDUAL FINAL BOOY WEIGHTS (g) ,, LIVER WEIGHTS (giI , AND RELATIVE LF/ER TO BODY WEIGHT OF MALE RATS Animal Number 625354 625365 625371 625376 625381 Final Body Wt. 396.1 459.6 447.3 432.9 381.8 GROUP IV Liver Wt. 16.383 18.313 17.859 19.168 14.135 Relative Liver to Body Wt. 0.041 0.040 0.040 0.044 0.037 Test Day 24 24 24 24 24 Animal Number 625356 625359 625360 625372 625374 Final Body Wt. 533.7 530.3 555.4 506.2 512.3 GROUP V Liver Wt. 20.498 22.901 23.312 21.287 18.378 Relative Liver to Body Wt. 0.038 0.043 0.042 0.042 0.036 Test Day 52 52 52 52 52 Animal Number 625353 625369 625373 625375 625377 Final Body Wt. 565.3 514.8 673.7 588.8 496.6 GROUP VI Liver Wt. 18.655 17.379 20.073 18.869 17.261 Relative Liver to Body Wt. 0.033 0.034 0.030 0.032 0.035 Test Day 94 94 94 94 94 -27- TSCACB1 Company Sanded. Does not contain H-24042: Biopersistelice Screening 10-Dose Oral Gavage,Study in Rats DuPont-2931 Supplement No. 1 H-2'1019 (POSITIVE ICONTROL) INDI\?TDUAL FINAL BODY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LB/ER TO BODY WEII3HT OF MALE RATS Animal Number 625322 625323 625336 625343 625349 Final Body Wfc. 329.0 298.0 318.1 294.0 293.7 GROUP I Liver Wt. 16.409 14.830 16.545 13.809 15.685 Relative Liver to Body Wt. 0.050 0.050 0.052 0.047 0.053 Test Day 5 5 5 5 5 Animal Number 625320 625330 625332 625341 625350 Final Body Wt. 312.1 310.1 318.3 325.6 284-.3 GROUP II Liver Wt. 18.845 16.343 17.528 18.254 15.709 Relative Liver to Body Wt. 0.060 0.053 0.055 0.056 0.055 Test Day 10 10 10 10 10 Animal Number 625321 625327 625342 625345 625348 Final Body Wt. 371.2 355.3 324.4 336.9 336.4 GROUP III Liver Wt. 22.322 22.259 20.718 24.403 20.541 Relative Liver to Body Wt. 0.060 0.063 0.064 0.072 0.061 Test Day 13 13 13 13 13 '2S' Company Sanitized. Does not contain TSCA CBi H-24042: Biopersistelice Screening 10-Dose Oral Gavage iStudy in Rats DuPont-2931 Supplement No. 1 H-2^1019 (POSITIVE CONTROL) IMDIVriDUAL FINAL BO DY WEIGHTS (g) ,, LIVER WEIGHTS (g ) , AND RELATIVB L:H/ER TO BODY WEIGHT OF MALE RATS Animal Number 625325 625328 625338 625340 625352 Final Body Wt. 381.3 423.9 339.9 419.0 375.3 GROUP IV Liver Wt. 21.479 22.100 18.018 23.131 21.835 Relative Liver to Body Wt. 0.056 0.052 0.053 0.055 0.058 Test Day 24 24 24 24 24 Animal Number 625324 625329 625333 625334 625344 Final Body Wt. 447.4 498.1 427.4 426.5 391.0 GROUP V Liver Wt. 20.900 20.911 20.508 19.144 17.666 Relative Liver to Body Wt. 0.047 0.042 0.048 0.045 0.045 Test Day 52 52 52 52 52 Animal Number 625326 625335 625346 626347 625351 Final Body Wt. 577.5 579.5 556.9 638.9 636.8 GROUP VI Liver Wt. 23.749 21.548 23.945 27.416 25.615 Relative Liver to Body Wt. 0.041 0.037 0.043 0.043 0.040 Test Day 94 94 94 94 94 -29- Company Sanitized, Does not contain TSCA CBI H-24042: Biopersistciice Screening 10-Dose Oral Gavage!Study in Rats DuPont-2931 Supplement No. 1 H-2.4020 (POSITIVE iCONTROL) INDI\IDUAL FINAL BC>DY WEIGHTS (g) , LIVER WEIGHTS (g ), AMD RELATIVE LTTOR TO BODY WEII3HT OP MALE RATS Animal Number 627756 627767 627771 627773 627777 Final Body Wt. 237.5 248.6 239.6 262.3 236.0 GROUP I Liver Wt. 15.360 19.033 16.968 17.629 16.268 Relative Liver to Body Wt. 0.065 0.077 0.071 0.067 0.069 Test Day 5 5 5 5 . 5 Animal Number 627768 627774 627776 627782 627785 Final Body Wt. 293.5 338.2 254.4 268.2 267.9 GROUP II Liver Wt. 22.536 27.332 18.821 20.172 20.857 Relative Liver to Body Wt. 0.077 0.081 0.074 0.075 0.078 Test Day 10 10 10 10 10 Animal Number 627758 627763 627764 627787 627788 Final Body Wt. 308.7 285.8 306.7 234.9 309.4 GROUP III Liver Wfc. 26.021 22.123 21.526 16.498 22.730 Relative Liver to Body Wt. 0.084 0.077 0.070 0.070 0.073 Test Day 13 13 13 13 13 .30- Company Sanitized. Does not contain TSCACBI H-24042: Biopersistence Screening 10-Dose Oral Gavag;e Study in Rats DuPont-2931 Supplement No. 1 H-2i.1020 (POSITIVE CONTROL) INDIVIDUAL FINAL BODY WEIGHTS (g),, LIVER WEIGHTS (g), AMD RELATIVE LI^/ER TO BODY WEIGHT OF MALE RATS Animal Mumber 627759 627761 627770 627779 627786 Final Body Wt. 380.1 359.4 379.8 406.2 326.1 GROUP IV Liver Wt. 22.707 19.579 19.540 24.858 16.410 Relative Liver to Body Wt. 0.060 0.054 0.051 0.061 0.050 Test Day 24 24 24 24 24 Animal Mumber 627757 627760 627765 627772 627781 Final Body Wt. 436.7 456.5 455.6 513.5 421.0 GROUP V Liver Wt. 16.899 18.240 16.505 20.225 16.695 Relative Liver to Body Wt. 0.039 0.040 0.036 0.039 0.040 Test Day 52 52 52 52 52 Animal Number 627762 627769 627775 627778 627783 Final Body Wt. 528.6 609.6 552.8 445.3 557.2 GROUP VI Liver Wt. 17.732 19.324 17.296 15.628 21.044 Relative Liver to Body Wt. 0.034 0.032 0.031 0.035 0.038 Test Day 94 94 94 94 94 -31- Company Sanitized. Doss not contain TSCA CB1 H-24042: Biopersisterice Screening 10-Dose Oral Gavage;Study in Rats DuPont-2931 Supplement No. 1 H-:24042 (TEST SUB,STANCE) INDI\ODUAL FINAL BOOY WEIGHTS (g), LIVER WEIGHTS (g), AND RELATIVE LB7ER TO BODY WEII3HT OP MALE RATS Animal Number 627295 627298 627300 627302 627311 Final Body Wt. 278.0 241.9 259.6 257.0 257.9 GROUP I Liver Wt. 18.201 13.763 16.695 16.611 16.097 Relative Liver to Body Wt. 0.065 0.057 0.064 0.065 0.062 Test Day 5 5 5 5 5 Animal number 627283 627289 627292 627293 627309 Final Body Wt. 264.4 277.6 308.6 274.2 306.6 GROUP II Liver Wt. 15.437 16.911 19.006 16.548 19.975 Relative Liver to Body Wt. 0.058 0.061 0.062 0.060 0.065 Test Day 10 10 10 10 10 Animal Number 627285 627288 627294 627301 627304 Final Body Wt. 314.3 316.5 292.6 343.6 321.4 GROUP III Liver Wt. 20.897 20.220 19.927 23.344 22.845 Relative Liver to Body Wt. 0.066 0.064 0.068 0.068 0.071 Test Day 13 13 13 13 13 -32- Company Sanitized. Does not contain TSCACBi H-24042: Biopersistelice Screening 10-Dose Oral Gavage iStudy in Rats DuPont-2931 Supplement No. 1 H-;}4042 (TEST SUBSTANCE) INDI\riDUAL FINAL BODY WEIGHTS (g),, LIVER WEIGHTS (g), AND RELATIVE LI^7ER TO BODY WEIGHT OF MALE RATS Animal Number 627279 627282 627286 627287 627299 Final Body Wt. 368.4 346.2 411.4 396.6 403.6 GROUP IV Liver Wt. 17.672 13.799 19.798 17.616 19.014 Relative Liver to Body Wt. 0.048 0.040 0.048 0.044 0.047 Test Day 24 24 24 24 24 Animal Number 627290 627296 627306 627307 627310 Final Body Wt. 438.4 411.4 497.7 527.6 525.6 GROUP V Liver Wt. 19.563 15.623 17.617 21.739 20.699 Relative Liver to Body Wt. 0.045 0.038 0.035 0.041 0.039 Test Day 52 52 52 52 52 Animal Number 627281 627297 627303 627305 627308 Final Body Wt. 574.3 531.5 512.5 743.2 598.6 GROUP VI Liver Wt. 18.730 18.196 16.634 27.873 19.317 Relative Liver to Body Wt. 0.033 0.034 0.032 0.038 0.032 Test Day 94 94 94 94 94 -33- Company Sanitized. Does not contain TSCA CBI H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2931 Supplement No. 1 APPENDIX B Individual Fluorine Levels -34- CaEspaay Sanitized. Does not contain TSCA CB1 Terms: Active % Active TERMS AND CALCULATIONS Mol Wt Active Formulation Dose % P in Active MolWtF Compound Calculations: Dose Active (mg/kg) The mg of fluorine containing compound administered per kg of animal body weight. = (% active/100) x Formulation Dose Dose Active (mmole/kg) The mmole of fluorine containing compound administered per kg of animal body weight = dose [mg/kg] / Mol Wt Active [mg/mmol] Dose F (mg/kg) The mg Fluorine administered per kg of animal body weight = (% F in active/100) x Dose Active [mg/kg] Dose F (mmol/kg) The mmole of fluorine administered per kg of animal body weight = Dose F [mg/kg] / Mol Wt F [mg/mmol] Molar Ratio (Active/P) The moles of fluorine containing compound per mole of fluorine = Dose Active [mmol/kg] / Dose P [mmol/kg] Formulation Dose Normalization Factor The formulation dose that would be required to administer the amount of active needed to achieve the normalized dose = (Normalized dose of Active [mmol/kg] / Dose Active [mmol/kg]) x Formulation Dose -35- Company Saait'szed. Does not contain TSCACBI H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2931 Supplement No. 1 TERMS AND CALCULATIONS Individual Animal Measurement: ppm F in liver The ppm fluoride measured in liver Individual Animal Calculations: ppm F in Liver minus Bkg 0.2 ppm ppm F in liver normalized to 0.1 mmol/kg Dose The ppm fluoride measured in liver minus the background fluoride measured in control animal liver. In this case the value was established at 0.2 ppm. The ppm fluoride minus background that would be expected if the active dose was 0.1 mmol/kg instead of the actual active dose. This assumes linearity between administered dose and liver fluorine levels, but is needed because different doses of active were used in the study. = (0.1 [mmol/kg] /Active dose [mmol/kg]) x (ppm F in liver minus background) /zmolar equivalents of active in liver The /xmolar [^mol/L] concentration of fluorine containing compound in liver based on the ppm fluorine in liver normalized to 0.1 mmol/kg active dose. This assumes that all liver fluorine is derived from the fluorine-containing component in the formulation. Note: 1 ppm - Img/L = (Normalized ppm [mg/L] fluorine in liver / Mol Wt F [mg/mmol]) x molar ratio active/F [mmol active/mmol F] x 1000 /imol/mmol 36- Compaq Sashed. Qoes not contain TSCA.CBS H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2931 Supplement No. 1 FACTORS INFLUENCING INTERPRETATION OF KINETIC ANALYSIS Considerations: - The data used for kinetic analysis was from a limited screen and extrapolation should be done cautiously. - Sample size is low - Analytical data used without validation - Steady-state not reached - Terminal phase may not be reached - Some compounds are mixtures of fluorinated compounds - Different active and formulation doses were used - Different vehicles were used to deliver formulations - Each compound may have very different potency for producing toxicity Assumptions: (May or may not be justified in all cases) - Liver fluorine concentrations are linear with respect to dose - Analytical method is appropriate for all types of compounds - Elimination kinetics can be determined based on total fluorine rather than on concentrations of individual components - Background Fluorine is 0.2 ppm - % F data is the % Fluorine of the active (Fluorine containing components) in the formulation) - Molecular weight is the molecular weight of the active component in the formulation Company SanitizetLDoesiiQtcsyitaJB-TSCA.CBi H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H-24019 DuPont-2931 Supplement No. 1 Given: Mol Wt. Active (g/mole): 497 Formulation Dose (mg/kg): 10 % Active (F Containing) in Formulation: 100 %F in Active: 65 Mol Wt.F(g/mol): 19 Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor: 10 0.020 49.7 DoseF(mmol/kg): 0.342 Molar Ratio (Active/F): 0.059 Dose F (mg/kg):_____6.5 Rat Number Test Day Sample ppmF in Liver Group II 625320 10 319.4 625330 10 355.4 625332 10 338.4 625341 10 305.0 625350 10 322.7 ppmF in Liver Minus Bkg 0.2 ppm 319.2 355.2 338.2 304.8 322.5 Group VI 625326 94 625335 94 625346 94 625347 94 625351 94 79.8 89.6 73.8 62.5 82.1 79.6 89.4 73.6 62.3 81.9 ppm F in Liver Normalized to 0.1 mmoles/kg Dose 1586.42 1765.34 1680.85 1514.86 1602.83 395.61 444.32 365.79 309.63 407.04 /(molar Equivalents of Active in Liver 4910.77 5464.62 5203.08 4689.23 4961.54 1224.62 1375.38 1132.31 958.46 1260.00 -38- Company Sanitized. Does not contain T3CA CB1 H-24042: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H-24020 DuPont-2931 Supplement No. 1 Given: Mol Wt. Active (g/mole): 426 Formulation Dose (mg/kg): 20 % Active (F Containing) in Formulation: 100 % F in Active: 69 MolWt.F(g/mol): 19 Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor: 20 0.047 42.6 DoseF(mmol/kg): 0.726 Molar Ratio (Active/F): 0.065 Dose F (mg/kg):_____13.8 Rat Number Test Day Sample ppmF in Liver Group II 627768 10 627774 10 627776 10 627782 10 627785 10 182.5 109.0 174.0 147.5 165.8 ppmF in Liver Minus Bkg 0.2 ppm 182.3 108.8 173.8 147.3 165.6 Group VI 627778 94 1.9 1.7 627783 94 1.2 1.0 627762 94 1.9 1.7 627775 94 1.6 1.4 627769 94 1.8 1.6 ppm F in Liver Normalized to 0.1 mmoles/kg Dose 388.30 231.74 370.19 313.75 352.73 3.62 2.13 3.62 2.98 3.41 fimolar Equivalents of Active in Liver 1321.01 788.41 1259.42 1067.39 1200.00 12.32 7.25 12.32 10.14 11.59 -39- Company Sanitized. Does not contain TBCA CBI 11204-D0o4s2e:OrBailopGearvsaigsteenScte yScrieneRnaiinsg SuppDleumpeonntt N2o9.31| - Data for H-24042 -- Give`nMo:l We. Active (g/mole): 531 % Fin Active: 75 Formulation Dose (mg/kg): 100 Mol Wi F (g/mol): 19 9% Active (F Containing) in Formulation: 100 CalcDuolsaeteAdctViavleue(sm:g/kg): 100 FDoorsmeulAacttiiovneD(omsmoelNeo/rkmea)l:ization Factor: 0.188 53.1 Dose F (mmol/kg): 3.947 Molar Ratio (Active/): 0.048 Dose F (mg/kg): 75 Test ppmF Rat Day in Number Sample _ Liver ppm Fin ppm F in Liver Liver MinusBkg ~~ Normalized to 0.1 mmoleskg 0.2 ppm Dose molar Equivalents of Activein Liver Group II 6278 10 1949 1947 10339 259.60 - 627289 10 2340 238 no 10 2141 2139 124.15 113.58 31173 285.20 6793 10 1821 621309 10 1936 1819 193.4 96.59 10270 24253 25787 Group VI 62781 9% 103 10.1 536 13.47 627297 m3 9% 94 114 122 12 120 595 637 14.93 1600 627305 94 149 627308 94 90 147 88 781 467 19.60 173 ---- _ o`CompanySanilizad. 009snotcontainTSCACBI